Zhong Xi Z, Dong Xian-Ping
Department of Physiology and Biophysics, Dalhousie University, Halifax, Nova Scotia, Canada.
Methods Cell Biol. 2015;126:197-215. doi: 10.1016/bs.mcb.2014.10.022. Epub 2015 Jan 14.
The physiology and functions of ion channels have been major topics of interest in biomedical research. Patch clamping is one of the most powerful techniques used in the study of ion channels and has been widely applied to the investigation of electrical properties of ion channels on the plasma membrane in a variety of cells. A number of ion channels have been found in intracellular lysosomal membranes. However, their properties had been difficult to study due to the lack of a direct patch-clamping methodology on lysosomal membranes. Past attempts to record lysosomal channels that were forced to express on the plasma membrane or reconstituted into lipid bilayers have largely generated inconclusive and conflicting results. Recently, a novel lysosome patch-clamping technique has been developed, making it possible to examine lysosomal channels under near physiological conditions. This chapter provides a detailed description of this technique, which has been successfully applied in several studies concerning lysosomal ion channels. This technique will expand our understanding of the nature of lysosomes and lysosome-related diseases.
离子通道的生理学和功能一直是生物医学研究中备受关注的主要课题。膜片钳技术是研究离子通道最强大的技术之一，已广泛应用于各种细胞中质膜上离子通道电特性的研究。在细胞内溶酶体膜中发现了许多离子通道。然而，由于缺乏对溶酶体膜进行直接膜片钳记录的方法，其特性一直难以研究。过去试图记录被迫在质膜上表达或重组到脂质双分子层中的溶酶体通道，大多产生了不确定和相互矛盾的结果。最近，一种新型的溶酶体膜片钳技术已经开发出来，使得在接近生理条件下研究溶酶体通道成为可能。本章详细描述了该技术，该技术已成功应用于几项关于溶酶体离子通道的研究中。这项技术将扩展我们对溶酶体本质和溶酶体相关疾病的理解。
